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原子力显微镜检测早期细胞凋亡过程中间充质干细胞的纳米形态和力学重建。

Nanomorphological and mechanical reconstruction of mesenchymal stem cells during early apoptosis detected by atomic force microscopy.

机构信息

Key Laboratory of Stomatology of State Ethnic Affairs Commission, Northwest Minzu University, Lanzhou 730030, China.

Key Laboratory of Mechanics on Disaster and Environment in Western China, The Ministry of Education of China, Lanzhou University, Lanzhou 730000, China.

出版信息

Biol Open. 2020 Mar 30;9(3):bio048108. doi: 10.1242/bio.048108.

Abstract

Stem cell apoptosis exists widely in embryonic development, tissue regeneration, repair, aging and pathophysiology of disease. The molecular mechanism of stem cell apoptosis has been extensively investigated. However, alterations in biomechanics and nanomorphology have rarely been studied. Therefore, an apoptosis model was established for bone marrow mesenchymal stem cells (BMSCs) and the reconstruction of the mechanical properties and nanomorphology of the cells were investigated in detail. Atomic force microscopy (AFM), scanning electron microscopy (SEM), laser scanning confocal microscopy (LSCM), flow cytometry and Cell Counting Kit-8 analysis were applied to assess the cellular elasticity modulus, geometry, nanomorphology, cell surface ultrastructure, biological viability and early apoptotic signals (phosphatidylserine, PS). The results indicated that the cellular elastic modulus and volume significantly decreased, whereas the cell surface roughness obviously increased during the first 3 h of cytochalasin B (CB) treatment. Moreover, these alterations preceded the exposure of biological apoptotic signal PS. These findings suggested that cellular mechanical damage is connected with the apoptosis of BMSCs, and the alterations in mechanics and nanomorphology may be a sensitive index to detect alterations in cell viability during apoptosis. The results contribute to further understanding of apoptosis from the perspective of cell mechanics.

摘要

干细胞凋亡广泛存在于胚胎发育、组织再生、修复、衰老和疾病的病理生理学过程中。干细胞凋亡的分子机制已得到广泛研究。然而,生物力学和纳米形态的改变却很少被研究。因此,我们建立了骨髓间充质干细胞(BMSCs)的凋亡模型,并详细研究了细胞力学特性和纳米形态的重建。原子力显微镜(AFM)、扫描电子显微镜(SEM)、激光共聚焦扫描显微镜(LSCM)、流式细胞术和细胞计数试剂盒-8 分析用于评估细胞弹性模量、细胞形态、纳米形态、细胞表面超微结构、生物活性和早期凋亡信号(磷脂酰丝氨酸,PS)。结果表明,在细胞松弛素 B(CB)处理的前 3 小时,细胞弹性模量和体积明显降低,而细胞表面粗糙度明显增加。此外,这些变化先于生物凋亡信号 PS 的暴露。这些发现表明细胞力学损伤与 BMSCs 的凋亡有关,力学和纳米形态的改变可能是检测细胞凋亡过程中细胞活力变化的敏感指标。这些结果有助于从细胞力学的角度进一步了解细胞凋亡。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3e1f/7132806/3829fbc7f075/biolopen-9-048108-g1.jpg

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